Abstract
Carotenes have attracted much attention in recent years for their biological function in processes such as photosynthesis. The characterization of carotenes is difficult, however, because they consist of only carbon and hydrogen atoms, without oxygen. In the present study, we systematically examined the chemical structures of more than 30 carotenes, including most of the carotenes found in phototrophic organisms, and observed their elution order using a Novapak C18 HPLC column with simple isocratic elution. The elution order of the carotenes was C30, C40,C45 then C50. The C40 carotenes with fewer conjugated double bonds (N) had longer retention times. With respect to the end groups, the carotenes eluted in the following order: φ, Ψ, ∈ then β end groups. Furthermore, absorption spectra in the HPLC eluent used were recorded with a photodiode-array detector. A greater N value was associated with a longer absorption maximum wavelength. Since the conjugated end groups (φ and β) influenced the absorption spectra and the non-conjugated end groups (Ψ and ∈) did not, the number of conjugated end groups (zero, one and two) was clearly distinguishable. Therefore, the chemical structures of carotenes can be easily determined by a combination of the HPLC retention times and the absorption spectra.
Similar content being viewed by others
References
Bianchi ML, Grein A, Julita P, Marnati MP and Spalla C (1970) Streptomyces mediolani (Arcamone et al.) emend. Bianchi et al. and its production of carotenoids. Z Allg Mikrobiol 10: 237–244
Britton G (1995) UV/visible spectroscopy. In: Britton G, Liaaen-Jensen S and Pfander H (eds) Carotenoids: Vol. 1B: Spectroscopy, pp 13–62. Birkhäuser, Basel
Britton G and Young AJ (1993) Methods for the isolation and analysis of carotenoids. In: Young A and Britton G (eds) Carotenoids in Photosynthesis, pp 409–457. Chapman & Hall, London
Buchecker R and Noack K (1995) Circular dichroism. In: Britton G, Liaaen-Jensen S and Pfander H (eds) Carotenoids: Vol 1B: Spectroscopy, pp 63–116. Birkhäuser, Basel
Craft NE (1992) Carotenoid reversed-phase high-performance liquid chromatography methods: Reference compendium. Methods Enzymol 213: 185–205
Davies BH and Köst H-P (1988) Carotenoids. In: Köst H-P (ed) CRC Handbook of Chromatography: Plant Pigments: Vol I, Fat-Soluble Pigments, pp 1–185. CRC Press, Boca Raton, Florida
Englert G (1995) NMR spectroscopy. In: Britton G, Liaaen-Jensen S and Pfander H (eds) Carotenoids: Vol. 1B: Spectroscopy, pp 147–260. Birkhäuser, Basel
Lancaster CRD and Michel H (1999) Refined crystal structures of reaction centres from Rhodopseudomonas viridis in complexes with the herbicide atrazine and two chiral atrazine derivatives also lead to a new model of the bound carotenoid. J Mol Biol 286: 883–898
Malhotra HG, Britton G and Goodwin TW (1970) A novel series of 1,2-dihydro carotenoids. Int J Vit Res 40: 315–322
Mayne ST (1996) Beta-carotene, carotenoids, and disease prevention in humans. FASEB J 10: 690–701
Nechushtai R, Eden A, Cohen Y and Klein J (1996) Introduction to Photosystem I: Reaction center function, composition and structure. In: Ort DR and Yocum CF (eds) Oxygenic Photosynthesis: The Light Reactions, pp 289–311. Kluwer Academic Publishers, Dordrecht, The Netherlands
Parker RS (1996) Absorption, metabolism and transport of carotenoids. FASEB J 10: 542–551
Pfander H (1992) Carotenoids: An overview. Methods Enzymol 213: 3–13
Pfander H and Riesen R (1995) Chromatography: Part IV High-performance liquid chromatography. In: Britton G, Liaaen-Jensen S and Pfander H (eds) Carotenoids: Vol 1A: Isolation and Analysis, pp 145–190. Birkhäuser, Basel
Sasa T, Suda S, Watanabe MM and Takaichi S (1992) A yellow marine Chlamydomonas: morphology and pigment composition, Plant Cell Physiol 33: 527–534
Satoh K (1996) Introduction to the Photosystem II reaction center: Isolation and biochemical and biophysical characterization. In: Ort DR and Yocum CF (eds) Oxygenic photosynthesis: The Light Reactions, pp 193–211. Kluwer Academic Publishers, Dordrecht, The Netherlands
Schiedt K and Liaaen-Jensen S (1995) Isolation and analysis. In: Britton G, Liaaen-Jensen S and Pfander H (eds) Carotenoids: Vol 1A: Isolation and Analysis, pp 81–108. Birkhäuser, Basel
Straub O (1987) Key to Carotenoids. Birkhäuser, Basel
Takaichi S (1993) Usefulness of field desorption mass spectrometry in determining molecular masses of carotenoids, natural carotenoid derivatives and their chemical derivatives. Org Mass Spectrom 28: 785–788
Takaichi S (1999) Carotenoids and carotenogenesis in anoxygenic photosynthetic bacteria. In: Frank HA, Young AJ, Britton G and Cogdell RJ (eds) The Photochemistry of Carotenoids, pp 39–69. Kluwer Academic Publishers, Dordrecht, The Netherlands
Takaichi S and Shimada K (1992) Characterization of carotenoids in photosynthetic bacteria. Methods Enzymol 213: 374–385
Takaichi S, Shimada K and Ishidsu J (1990) Carotenoids from the aerobic photosynthetic bacterium, Erythrobacter longus: β-carotene and its hydroxyl derivatives. Arch Microbiol 153: 118–122
Takaichi S, Tsuji K, Matsuura K and Shimada K (1995a) A monocyclic carotenoid glucoside ester is a major carotenoid in the green filamentous bacterium Chloroflexus aurantiacus. Plant Cell Physiol 36: 773–778
Takaichi S, Yazawa H and Yamamoto Y (1995b) Carotenoids of the fruiting gliding myxobacterium, Myxococcus sp. MY-18, isolated from lake sediment: Accumulation of phytoene and keto-myxocoxanthin glucoside ester. Biosci Biotechnol Biochem 59: 464–468
Takaichi S, Sandmann G, Schnurr G, Satomi Y, Suzuki A and Misawa N (1996) The carotenoid 7,8-dihydro-Ψ end group can be cyclized by the lycopene cyclases from the bacterium Erwinia uredovora and the higher plant Capsicum annuum. Eur J Biochem 241: 291–296
Takaichi S, Wang Z-Y, Umetsu M, Nozawa T, Shimada K and Madigan MT (1997a) New carotenoids from the thermophilic green sulfur bacterium Chlorobium tepidum: 1′,2′-dihydro-γ-carotene, 1′,2′-dihydrochlorobactene, and OH-chlorobactene glucoside ester, and the carotenoid composition of different strains. Arch Microbiol 168: 270–276
Takaichi S, Inoue K, Akaike M, Kobayashi M, Oh-oka H and Madigan MT (1997b) The major carotenoid in all known species of heliobacteria is the C30 carotenoid 4,4′-diaponeurosporene, not neurosporene. Arch Microbiol 168: 277–281
Vertès AA, Asai Y, Inui M, Kobayashi M, Kurusu Y and Yukawa H (1994) Transposon mutagenesis of coryneform bacteria. Mol Gen Genet 245: 397–405
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Takaichi, S. Characterization of carotenes in a combination of a C18 HPLC column with isocratic elution and absorption spectra with a photodiode-array detector. Photosynthesis Research 65, 93–99 (2000). https://doi.org/10.1023/A:1006445503030
Issue Date:
DOI: https://doi.org/10.1023/A:1006445503030